紧密连接蛋白-5（Claudin-5）及血管内皮钙粘蛋白（VE-cadherin)在海绵状血管畸形出血风险中的作用及机制研究

脑和脊髓海绵状血管畸形是先天性的出血性血管疾病，不同患者的出血风险及严重程度差异很大，尚未发现特异性的血生物标志物。研究发现，内皮细胞连接缺陷与病变出血风险及严重程度相关，但机制尚不清楚。紧密连接蛋白Claudin-5和钙粘着蛋白VE-cadherin是维持内皮细胞间连接最重要的两种蛋白，我们推测可能是该病潜在的特异性生物标志物，且可以在周围血中检测。本研究将:1)利用大样本患者队列对上述蛋白与出血频次及严重程度进行关联分析，验证其作为该病特异性血生物标记物的可行性及有效性;2)在体外细胞实验中，观察内皮细胞经上述蛋白单克隆抗体干预后的表型变化;3)对ccm突变斑马鱼模型敲低上述蛋白表达水平，在体内实验中观察对病变表型及内皮细胞的影响。本研究将阐明Claudin-5和VE-cadherin在海绵状血管畸形发生发展及出血风险中的作用及机制，为该病的机制研究和诊疗方案奠定理论基础。

Cavernous malformation in the central nervous system is a congenital hemorrhagic vascular disease. The hemorrhage risk and clinical severity of different patients vary widely, and there are still no accepted specific blood biomarkers for predicting disease progression. Previous studies have found that endothelial cell junction defects play a key role in the mechanism and clinical manifestations of cavernous malformation, while the mechanism remains unclear. Claudin-5 and VE-cadherin are the two most important proteins that maintain intercellular endothelial junctions. And they can be detected in the blood and we infer that they are potential biomarkers for cavernous malformation. Our group has one of the largest case series of cerebral and spinal cord vascular malformations in the world. We propose to use a large sample of patients to correlate the above proteins with clinical processes to verify the feasibility of being a specific biomarker for cavernous malformation; In vitro cell experiments, we will observe the phenotypic changes of endothelial cells after intervention with the above-mentioned protein monoclonal antibody; And morpholine antisense oligonucleotide technology will be used in the ccm mutant zebrafish model to knock down the above protein expression. By analyzing its effect on the phenotype, endothelial cell migration and cell number in vivo and in vitro, we try to elucidate the mechanism of Claudin-5 and VE-cadherin that affect the development and hemorrhage events of cavernous malformation in the central nervous system, which will provide theoretical basis for the mechanism study and clinical application of cavernous malformation.